| Objective:(1) To employ the new method: loop-mediated isothermal amplification (LAMP) and PCR to test the HBV-DNA. During this process, We optimized the reacting condition of the LAMP,and made comparison of the specificity and sensitivity between these two methods. (2) To evaluate the anti-HBV function of Supplement Qi for Activating Blood Circulation TCM on 2.2.15 cells line. Besides we evaluated the inhibitory effects of the herbs by using LAMP method.Methods:(1) Collection of the samples: the sera samples were collected from the patients ingfected with HBV, and HBV-DNA was extracted from the samples as template to amplify DNA using LAMP method. Based on the above works, we optimized the reaction condition for LAMP and establisheda novel method for amplification of the HBV-DNA.(2) Confirmation of the sensitivity of the LAMP: we detected the same samples by the two methods-LAMP and PCR simultaneously, and compared the sensitivity of the LAMP with that of PCR.(3) Confirmation of the specificity of the LAMP: we detected the HBV-positive sera samples , the DHBV-positive sera samples and negative sera samples by LAMP, and observed the occurrence of the false positive results in the test.(4) Inhibitory function of Supplement Qi for Activating Blood herbs on the HBV by using the 2.2.15 cell model: based on the 2.2.15 cell line, we detected the cytotoxic effects of the herbs andthe Inhibitory effects of the herbs on HBsAg, HBeAg secretion,evaluated the herbs of the inhibitionof HBV on the cellular level with the therapeutic index(TI) in vitro.(5) Establishment of the LAMP method: employ LAMP methods on 2.2.15 cell lines which has been cultured under the water extraction of the herbs, to detect the HBV DNA figures and evaluate the inhibitory function on HBV of the herbs. On all the above to establish a new detecting method for KBV-detection.Results:(1) Succeed in amplification of the HBV-DNA by using the LAMP method, and modify the... |
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